Department of Veterinary Microbiology and Pathology, College of Veterinary Medicine, Washington State University, Pullman, WA 99164-7040, United States.
J Microbiol Methods. 2010 Jul;82(1):36-41. doi: 10.1016/j.mimet.2010.03.017. Epub 2010 Mar 31.
A microarray was developed to simultaneously screen Escherichia coli and Salmonella enterica for multiple genetic traits. The final array included 203 60-mer oligonucleotide probes, including 117 for resistance genes, 16 for virulence genes, 25 for replicon markers, and 45 other markers. Validity of the array was tested by assessing inter-laboratory agreement among four collaborating groups using a blinded study design. Internal validation indicated that the assay was reliable (area under the receiver-operator characteristic curve=0.97). Inter-laboratory agreement, however, was poor when estimated using the intraclass correlation coefficient, which ranged from 0.27 (95% confidence interval 0.24, 0.29) to 0.29 (0.23, 0.34). These findings suggest that extensive testing and procedure standardization will be needed before bacterial genotyping arrays can be readily shared between laboratories.
研制出一种微阵列,用于同时对大肠杆菌和沙门氏菌进行多种遗传特征的筛选。最终的微阵列包括 203 个 60 碱基寡核苷酸探针,包括 117 个耐药基因、16 个毒力基因、25 个复制子标记和 45 个其他标记。通过使用盲法研究设计评估四个合作小组之间的实验室间一致性来测试微阵列的有效性。内部验证表明该检测方法是可靠的(接受者操作特性曲线下的面积=0.97)。然而,使用组内相关系数估计的实验室间一致性较差,范围从 0.27(95%置信区间 0.24,0.29)到 0.29(0.23,0.34)。这些发现表明,在细菌基因分型微阵列能够在实验室之间方便地共享之前,需要进行广泛的测试和程序标准化。